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10-year-old India Opal Buloni has just moved to the fictional small town of Naomi, Florida with her father, a preacher.

While in the Winn-Dixie supermarket, she encounters a scruffy Berger Picard that is wreaking havoc.[2] Opal (not wanting the store manager to send the dog to the pound) claims that it is her dog and names it "Winn-Dixie". Winn-Dixie becomes friends with everyone he encounters, and so Opal makes some new friends in the process. She also rekindles her relationship with her father, and learns ten things about her mother, who abandoned them seven years ago. Opal describes the preacher as a turtle, always sticking his head into his turtle shell, and never wanting to come out into the real world. This is most likely because of how sad he is about her mother, with whom he is still in love.

One of the people Opal meets is Miss Franny Block, a kind and somewhat eccentric elder librarian, who tells her many great stories, including one involving a bear. Opal also meets Gloria Dump, a blind recovering alcoholic with a tree in her backyard that has beer bottles hanging from it. She calls it a 'mistake tree' and the bottles represent the ghosts of all the things she has done wrong. One day, fed up with Winn-Dixie, the landlord of the Bulonis' trailer park, Mr. Alfred, orders the preacher to get rid of the dog. The preacher calls the animal pound to take Winn-Dixie away, but Opal begs her father to keep her dog. Unable to see his daughter this upset, the preacher tells the pound to return Winn-Dixie, claiming that he is not the same dog he called about.

Opal gets a job at Gertrude's Pets and befriends a worker there, Otis, a shy ex-convict with a passion for music. She also meets a young girl named Sweetie Pie Thomas, who is eager to get a dog like Winn-Dixie. Later, a thunderstorm comes and Winn-Dixie, being pathologically afraid of thunderstorms, runs away. While Opal looks for him, her father wants to give up and she blames him for the loss of her mother and Winn-Dixie running away. But her father explains that he tried very hard to look for her mother. He then admits that he believes that she is never coming back. Later they go back to a party and Otis starts to sing a song on his guitar. Winn-Dixie is heard outside howling along to the song. Everyone, while singing, lets him in and welcomes him back.

The film was shot on location in Napoleonville, Louisiana, USA. To make sure both dogs got on well with AnnaSophia Robb, who played Opal, she was brought in early to get acquainted with the dogs and give them treats. By the time shooting started, they considered her a "safe" area.[1] Winn-Dixie was played by multiple Picardy Shepherds, a rare breed from France. The DVD extra "Diamond in the Ruff" shows the two principal dogs, Scott and Lyco, but producer Trevor Albert mentions at 18:00 and 40:26 in the DVD feature commentary that, in all, four dogs were used. At 00:36 in AnnaSophia Robb's commentary "Meet Winn-Dixie" she mentions that the stunt dog Tasha jumped over the flour. The film's mouse was played by a rat. The choice was made carefully because while a mouse would have been preferable, rats are much easier to train.

Director Wayne Wang wanted to use Picardy Shepherds because he thought they looked similar to the depiction of Winn-Dixie on the book cover and would appear familiar to its readers. Dogs were brought from France when none were available in the U.S. (08:55 in the DVD commentary). The film, like the book, is set in Naomi, Florida, even though it was filmed in Louisiana. Consequently, the police car and uniform emblems depict the state of Florida rather than the state of Louisiana. The bunny that Ottis hands Opal (at around 56 mins) is a Netherlands Dwarf. They only get to be between 6 and 8 inches long. The last name of Opal and her dad, Buloni (mentioned in the dialog at 49:50 in the lunch meat joke), is shown on a sign to the left of the trailer door at 22:37 and elsewhere in the feature. In the scene from 10:45 to 11:03 where Winn-Dixie first arrives at the trailer, only the bottom edge of the sign is visible in the full frame version at 10:53 (when seen on a computer without "overscan" cropping) while in the wide screen version that edge is cropped from view.

This was Jeff Daniels and Harland Williams' second movie together; both had previously appeared in Dumb and Dumber (1994). The rat used for the beginning of the church scene is male, it then switches to being a female rat, and back again between shots. During the scene where the animals get loose, the same event is used twice. When Opal picks up the black and white rabbit to place in its pen, it is shown once as a close up of her [at 33:41 from behind] and once from a distance [at 33:58 frontal]. (This can be clearly seen in the full frame version, but not in the wide screen version at 33:41 where the wide screen cropping of the spherical 35 mm frames crop out the body of the rabbit. Consequently, it is a goof in re-mastering the full frame version from the spherical 35 mm negatives rather than a goof in the wide screen film. Of course, it could be argued that the rabbit got out again and needed to be put into the pen a second time.) In the scene where Opal brings Winn-Dixie into the church, when she gets up from her seat to get Winn-Dixie, there is a Bible and a purse on the chair beside her. After she returns from getting Winn-Dixie, there is somebody sitting in the seat beside her holding the Bible while the purse is on the floor between the two chairs. Opal's hair length varies throughout the movie, from a short blunt cut, to a longer, and uneven cut, and then back to short. When it starts raining, at the start of the garden party, all of the lights strung around start to flicker and go out. Except when you see everyone standing at the house, the lights to the right can be seen still on. Opal's hair changes from straight and flat to styled and blow-dried during the baseball game and party set-up scenes. The "mouse" Winn Dixie catches during the church service is a rat.

The film received mixed reviews from critics. Rotten Tomatoes gives the film a score of 55% based on 121 reviews, with an average rating of 5.8/10. The site's consensus was: "An old-fashioned, if bland, adaptation of Kate DiCamillo's novel."[3] At Metacritic, the film holds a score of 54% based on 27 reviews, indicating "mixed or average reviews".[4]